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A rate turbidimetric immunoassay for theophylline using biotin-avidin system
Insights
A new biotin-avidin immunoassay accurately measures serum theophylline levels. This assay uses turbidity to quantify theophylline, showing good correlation with existing methods.
Area of Science:
- Clinical Chemistry
- Biochemistry
- Immunology
Background:
- Serum theophylline monitoring is crucial for managing respiratory diseases.
- Existing assays may have limitations in speed or accessibility.
Purpose of the Study:
- To develop and validate a novel biotin-avidin based rate turbidimetric homogeneous immunoassay for serum theophylline determination.
Main Methods:
- Utilized an avidin-biotin-labeled theophylline conjugate and a monoclonal antibody.
- Monitored the rate of turbidity change on the Beckman Synchron CX 4 System.
- Quantified theophylline by measuring inhibition of antibody-conjugate complexation.
Main Results:
- Achieved a measuring range of 0-40 micrograms/ml for serum theophylline.
- Demonstrated excellent precision with within-run CV < 3.7% and between-run CV 5.2-9.2%.
- Showed high correlation (r = 0.9866) with a fluorescence polarization immunoassay.
Conclusions:
- The developed assay is a reliable and precise method for serum theophylline quantification.
- This turbidimetric immunoassay offers a viable alternative for clinical laboratories.
Abstract:
We describe a biotin-avidin based rate turbidimetric homogeneous immunoassay for the determination of serum theophylline with a measuring range of 0-40 micrograms/ml. The assay features an avidin-biotin-labeled theophylline conjugate and a monoclonal antibody. The rate of change of turbidity caused by the antibody-conjugate complexing was monitored on the Beckman Synchron CX 4 System at 340 nm and 37 degrees C. Theophylline in a sample inhibited the complexation, and the extent of inhibition allowed the quantitation of theophylline in the sample. The within-run coefficient of variation (CV) was < 3.7% and the between-run CV was 5.2-9.2% depending on the theophylline concentration. Linear regression analysis showed a good correlation with an established fluorescence polarization immunoassay (r = 0.9866, n = 94).
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